9h in distilled water (pHp5.3), citric acid solution (0.1%,. pHp2.6) or sodium bicarbonate solution (0.07%, pHp8.4). The seed to solution ratio was 1:3 (w/v).
Eur Food Res Technol (2000) 210 : 340–345
Q Springer-Verlag 2000
ORIGINAL PAPER
Juana Frias 7 Concepción Vidal-Valverde Cristina Sotomayor 7 Concepción Diaz-Pollan Gloria Urbano
Influence of processing on available carbohydrate content and antinutritional factors of chickpeas Received: 22 April 1999 / Revised version: 28 June 1999
Abstract The effect of chickpea processing (soaking, soaking plus cooking and dry heating) on the content of available carbohydrate (monosaccharides, disaccharides and starch) and antinutritional factors (a-galactosides and trypsin inhibitor activity) was studied. Soaking produced a reduction in available carbohydrates (19–20%) and a-galactosides (16–27%), and either did not affect or caused a slight reduction of trypsin inhibitor activity (TIA) in chickpeas. Soaking plus cooking brought about a larger decrease in available carbohydrates (23–24%) and a-galactosides (45–58%), and completely eliminated TIA. Dry heating caused a 24% reduction in available carbohydrates, a 46% decrease in a-galactosides and a 27% decrease in TIA. Overall, cooking the presoaked seeds in water, acidic or basic solution seems to be adequate to obtain a chickpea flour with a large reduction in antinutritional factors (a-galactosides and TIA) and also a high level of available carbohydrate content. Key words Carbohydrates 7 Chickpeas 7 Starch 7 a-Galactosides 7 Trypsin inhibitor activity
Introduction Chickpeas (Cicer arietinum L.) are the fifth most important legume in the world on the basis of total grain production after soybeans, peanuts, beans and peas [1]. They are the most widely consumed legume throughout Spain and are especially popular in Andalusia [2].
J. Frias 7 C. Vidal-Valverde (Y) 7 C. Sotomayor C. Diaz-Pollan Instituto de Fermentaciones Industriales (CSIC), Juan de la Cierva 3, E-28006 Madrid, Spain e-mail: ificv126ifi.csic.es G. Urbano Departamento de Fisiologia, Instituto de Nutricion, Facultad Farmacia, Campus Universitario de Cartuja s/n, E-18071 Granada, Spain
Chickpeas are a good source of protein (12.6–30.5%) [3], and exhibit higher true digestibility, biological value and net protein utilisation than cowpeas and mung beans [4], higher net protein utilisation than soybeans, peas, beans and lentils [5], and higher PER than soybeans, faba beans, pigeon peas, black gram and mung beans [6]. Chickpea seeds contain 54.4% to 70.9% of total carbohydrates, of which the major proportion is starch (37.2–50.8%) [1]. Although chickpeas are considered to be one of the most nutritious pulses [3], they contain several antinutritional factors that could limit their consumption and the nutritive utilisation of their proteins [7]. Among these are the flatulence-causing a-galactosides [8–10], which have been related to the excessive accumulation of gas in the intestine [11, 12]. Consequently, the presence of these sugars in chickpea seeds is one of the major constraints in their full utilisation as human food. Trypsin inhibitors are also present in chickpea seeds and they are capable of binding to the trypsin enzyme, thus inhibiting its activity, interfering with the digestion of proteins and resulting in an increased pancreatic secretion and hypertrophy of the pancreas [13, 14]. The removal of undesirable components is therefore essential to improve the nutritional quality of legumes and effectively utilise their full potential as human food. Simple and inexpensive processing techniques are an effective method of achieving desirable changes in the seed composition and improving palatability. Various authors have reported that soaking and heat treatments improve the quality of legumes. Soaking removes some antinutritional compounds, which can be partly or totally solubilised and eliminated with the discarded soaking solution. At the same time, some metabolic reactions take place during the soaking procedure, affecting the content and composition of the seeds [10]. Heat treatments generally inactivate heatsensitive factors such as trypsin inhibitors and remove volatile compounds. The cooking water may be discarded and some other soluble compounds removed. In
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many instances, the use of only one method may not result in the desired removal of antinutritional compounds and the combination of two or more methods is required. Although some information has been found concerning the chemical composition of chickpeas, not much work has been carried out on the effect of simple processes, or their combination, on the modification of the nutritional and antinutritional composition of chickpeas. The aim of this work was to evaluate the effect of simple treatments on the content of available carbohydrate and on some antinutritional factors of chickpeas in order to obtain chickpea flours with high nutritive value, which may be utilised for human consumption as part of the diet or in the fortification of foods.
Materials and methods Samples. Raw, dry chickpeas (R) (C. arietinum L. cv. Blanco lechoso) were grown in Andalusia (southern Spain). The seeds were subjected to seven different treatments: S p soaking in distilled water; SA p soaking in acidic medium; SB p soaking in basic medium; SC p S c cooking; SAC p SA c cooking; SBC p SB c cooking; H p dry heating. Soaking. Chickpea seeds were soaked at room temperature for 9 h in distilled water (pHp5.3), citric acid solution (0.1%, pHp2.6) or sodium bicarbonate solution (0.07%, pHp8.4). The seed to solution ratio was 1 : 3 (w/v). The soaking liquid was drained off, and the seeds were blended and lyophilised. Cooking. Soaked chickpeas seeds were cooked by boiling in distilled water for 35 min at a seed to water ratio of 1 : 6.67 (w/v). The cooking water was drained off, and the seeds were crushed and lyophilised. Dry heating. Raw ground chickpeas were dry heated under pressure at 120 7C and 1 atm for 15 min. Determination of monosaccharides, disaccharides and a-galactosides. The analysis of these compounds was carried out by highperformance liquid chromatography according to Frias et al. [15]. Starch determination. Starch was analysed from the residue obtained after soluble carbohydrate extraction using a procedure based on the total enzymatic digestion to glucose [16]. Glucose content was measured by the method of glucose-oxidase peroxidase [17]. The starch content was calculated by multiplying the resulting glucose content by 0.9. Trypsin inhibitor activity. Extraction of trypsin inhibitors was performed according to the method of Kakade et al. [18] modified by Valdebouze et al. [19]. Statistical methods. Multifactor analysis of variance was applied to the data using Statgraphic Statistical Graphics 5.0 System Software (Statistical Graphics Corporation, Rockville, Md., USA).
Results and discussion The effects of soaking in water, citric acid and sodium bicarbonate, soaking in those solutions plus cooking in
water, and dry heating at 120 7C at 1 atm for 15 min on the content of available carbohydrate and antinutritional factors [a-galactosides and trypsin inhibitor activity (TIA)] of chickpeas are shown in Tables 1 to 3. Starch content decreased by 20–21% after soaking. The type of solution employed did not affect this reduction (Table 1). The effect of soaking on the starch content is controversial because of the different results found in the literature. Chavan et al. [3], Jood et al. [20, 21], Ologhobo and Fetuda [22] and Vidal-Valverde et al. [23] reported that during soaking of Vicia faba, C. arietinum, Cajanus cajan, Phaseolus vulgaris and Phaseolus mungo the starch content decreases between 6% and 16%. These differences might be due to differences in starch structure, seed size and membrane permeability, since these factors may contribute to the different solubilisations of starch during soaking. In faba beans [23] and kidney beans [24] the pH during soaking did not significantly affect the starch content, whilst Jood et al. [20], working with V. faba, C. arietinum, P. vulgaris, P. mungo and Ca. cajan found a large starch decrease when soaking was carried out in basic solution compared with water. Sathe et al. [25] reported for P. mungo that soaking at acidic and basic pH caused a reduction in the viscosity possibly because of a larger starch breakdown giving rise to more sugar leaching. On the other hand, Frias [26] reported that soaking lentils in acidic and basic solution caused an increase in starch content compared with water soaking. These different results show that changes in starch content during soaking seem to depend not only on the type of solution employed but also on the legume investigated. Soaking plus cooking caused a decrease in starch content and reductions of 21–22% were also obtained, irrespective of the previous soaking solution employed (Table 1). Information in the literature about the effect of soaking plus cooking on the starch content of chickpeas is scarce. Various authors have reported that during soaking and cooking the decrease in starch content may be due not only to the type of legume and its physical characteristics [3, 21–23, 26–28], but also to whether or not a previous treatment has been carried out [21, 23, 28]. Our results indicate no significant differences in starch content between the soaking treatment and the soaking plus cooking treatment. Dry heating caused larger losses in starch content than those observed after soaking and cooking and a 26% reduction was obtained. These larger losses are probably caused by the high temperature and pressure used during dry heating, which could modify the starch structure differently compared with the soaking and cooking treatment. These results are in accordance with those reported by Jood et al. [20] for V. faba, by Ologhobo and Futuda [22] for P. lunatus, and by Jood et al. [21] for C. arietinum and Phaseolus. Kataria et al. [28] showed that dry heating of soaked Phaseolus caused larger starch reductions than in non-soaked Phaseolus. Fructose and sucrose did not show significant changes (P^0.05) after soaking compared with raw
342 Table 1 Effect of processing on available carbohydrate content of chickpeas. Values are the mean of four determinations B standard deviation. The same superscript in the same column means no significant differences (P^0.05) Cicer arietinum
Starch g/100 g (d.m.)
Unprocessed 52.56B1.66 Water soaking 41.53B1.16 a Citric acid soaking 42.04B1.07 a Sodium bicarbonate soaking 41.81B2.63 a Water soaking c cooking 40.74B2.15 a Citric acid soaking c cooking 40.99B1.70 a Sodium bicarbonate soaking c cooking 41.64B2.77 a Dry heating 38.95B1.43
Fructose Reduc- g/100 g (d.m.) tion (%) 21 20 20 22 22 21 26
chickpeas (Table 1). Cooking the soaked seeds brought about a large decrease in both available sugars: fructose showed a reduction between 42% and 83% and sucrose between 45% and 50%. These results agree with those published by Iyengar and Kulkarni [29] for lima beans, Iyer et al. [30] for Phaseolus and Jood et al. [20, 21] for different cultivars of chickpeas; these authors found larger losses after cooking compared with the soaking process. Taking into account the fact that the total available carbohydrate is the sum of starch, sucrose and fructose (Table 1), it can be said that the three soaking treatments carried out with chickpeas resulted in similar reductions of 19–20%. Similarly, with the three cooking treatments and dry heating a reduction of 23–24% occurs. These results show that when soaking or heat treatments are carried out on chickpeas, the total available carbohydrate content, as a whole, can still contribute very much to the energetic intake of the diet, although individual available carbohydrates can suffer from the effect of processing in different ways. Table 2 shows the content of raffinose, ciceritol, stachyose and total a-galactosides in raw and processed chickpeas. In raw seeds, ciceritol was present in the largest amount (2.7%) followed by stachyose (1.7%) and raffinose (0.5%), values which are in agreement with the range found in the literature [21, 31–35]. As a consequence of soaking, the a-galactoside content suffered a decrease that was sharper when soaking was carried out in water (a decrease of 27%) than either in acidic or basic medium (17% and 16% reductions, respectively) (Table 2). The individual a-galactosides behaved differently depending on the type of solution, raffinose suffered a decrease of 20–22% in chickpeas soaked in water and basic solution but only a 7% decrease after soaking in acidic solution. Ciceritol decreased by 30–32% after soaking in acidic and basic solution and by 40% when chickpeas were soaked in water. Stachyose, however, showed only a slight decrease (7%) in water-soaked chickpeas whilst when soaking was carried out in basic solution it suffered a fair but significant increase (P^0.05).
0.12B0.02 ab 0.14B0.03 a 0.13B0.02 a 0.11B0.01 b 0.07B0.01 0.02B0.01 0.04B0.02 0.16B0.01
Sucrose Reduc- g/100 g (d.m.) tion (%)
P P P 42 83 67 P
Total available carbohydrates Reduc- g/100 g (d.m.) tion (%)
3.53B0.42 a 3.29B0.35 a 7 3.49B0.25 a 1 3.35B0.22 a 5 1.93B0.08 b 45 1.76B0.07 b 50 1.78B0.09 b 50 3.40B0.32 a 4
56.21 44.81 45.66 45.27 42.74 42.77 43.46 42.51
Reduction (%) 20 19 19 24 24 23 24
These results are in agreement with the data found in the literature for different legumes and, as a consequence of soaking, a significant reduction of a-galactosides is achieved when the soaking solutions are removed. The decrease obtained depends not only on the type of legume and soaking solution but also on the process time and temperature [21, 23, 29, 30, 36–38]. The decrease obtained during the soaking process in total carbohydrates and especially in a-galactosides may be partly due to the solubilisation and leaching of these compounds to the soaking solution and also to some metabolic changes which take place during soaking, since the a-galactosides, which are storage products, can be metabolised. Vidal-Valverde et al. [10] observed this phenomenon during lentil soaking. Greater reduction in total a-galactoside content was achieved after the soaking plus cooking treatment (Table 2), and the reduction obtained ranged between 45% and 58%. In these treatments all individual a-galactosides suffered a remarkable decrease. According to information from the literature, the elimination of a-galactosides in legumes after soaking plus cooking seems to be connected with the removal of the soaking and cooking solution. When the processing liquids are not discarded, a increase or no marked decreases in a-galactoside content is observed [20–22, 25, 28, 39, 40] whilst removing the processing solutions causes a large reduction in a-galactosides [10, 23, 29, 30, 34]. Dry heating also caused a large a-galactoside decrease and a 46% reduction was obtained (Table 2). Raffinose and stachyose underwent decreases of 57% and 58% respectively, and a loss of 37% was observed for ciceritol, results which are in agreement with those presented by Jood et al. [20], who found that dry heating caused a 73% reduction of total a-galactosides in chickpeas. a-galactosides are considered as antinutritional factors because of their flatus production and discomfort after consumption of legumes [9], and they can also be associated with a low food intake in animal experiments. Nestares et al. [41] fed rats with the same raw and processed chickpeas used in the present paper and
343 Table 2 Effect of processing on the a-galactoside content of chickpeas. Values are the mean of four determinations B standard deviation. The same superscript in the same column means no significant differences (P^0.05) C. arietinum
Raffinose g/100 g (d.m.)
Unprocessed 0.46B0.04 a Water soaking 0.36B0.05 b Citric acid soaking 0.43B0.03 a Sodium bicarbonate soaking 0.37B0.03 b Water soaking c cooking 0.17B0.02 Citric acid soaking c cooking 0.13B0.01 c Sodium bicarbonate soaking c cooking 0.11B0.02 c Dry heating 0.20B0.02
Ciceritol Reduc- g/100 g (d.m.) tion (%) 22 7 20 63 72 76 57
they reported that the food intake, expressed as grams of diet per 100 g of body weight, was significantly greater for all processed diets than for raw chickpeas, which could be related to the decrease in a-galactosides obtained after processing. Table 3 shows changes in the content of trypsin inhibitor activity (TIA) of chickpeas during processing. For raw chickpeas a TIA of 10.4 units/mg d.m. was obtained, a content that is higher than that for peas, lentils and faba beans [5, 42–44]. Soni et al. [45] reported that trypsin inhibitor levels in chickpeas were 66% of that for soya bean. Soaking chickpeas in distilled water modified slightly TIA and a decrease of only 12% was found, whilst soaking in citric acid and sodium bicarbonate did not affect TIA content (Table 4). Information found in the literature about the effect of soaking on TIA of chickpeas is very scarce. Hamza et al. [46] observed that soaking of chickpeas, faba beans and lentils caused a decrease on electrophoretic bands of TIA. Vidal-Valverde et al. [47] reported that soaking faba beans in either water or sodium bicarbonate led to a decrease of 19% and 24% respectively, whilst soaking in citric acid did not influence the activity of trypsin inhibitors. Sharma and Sehgal [48] also reported that TIA of faba beans was not modified after 12 h of soaking in water. Table 3 Effect of processing on trypsin inhibitor activity (TIA) of chickpeas; ND p not detected (less than 0.5 TIU/mg). Values are the mean of four determinations B standard deviation C. arietinum
Unprocessed Water soaking Citric acid soaking Sodium bicarbonate soaking Water soaking c cooking Citric acid soaking c cooking Sodium bicarbonate soaking c cooking Dry heating
TIA Units/mg (d.m.)
Reduction (%)
10.43B0.77 a 9.20B0.67 10.47B0.46 a 10.77B0.75 a ND ND ND
12 P P 100 100 100
7.60B0.50
27
2.70B0.18 1.61B0.08 1.83B0.09 a 1.89B0.15 a 1.82B0.13 a 1.48B0.08 1.30B0.04 1.69B0.10
Stachyose Reduc- g/100 g (d.m.) tion (%) 40 32 30 33 45 52 37
1.68B0.16 ab 1.57B0.10 a 1.75B0.09 bc 1.82B0.16 c 0.68B0.09 d 0.66B0.09 d 0.60B0.06 d 0.71B0.11 d
Total a-galactosides Reduc- g/100 g (d.m.) tion (%) 7 P P 60 61 64 58
4.84 3.53 4.01 4.08 2.67 2.27 2.01 2.60
Reduction (%) 27 17 16 45 53 58 46
In Phaseolus, Sathe et al. [25] reported losses of 3.3% and 13.3% after soaking the seeds in water or basic solution. In soaked lentils, Batra et al. [49] observed TIA losses of 58–66%, whilst Vidal-Valverde et al. [50] reported losses of only 4–11% depending on the type of soaking solution employed. These results indicate that the effect of soaking on the activity of trypsin inhibitors depends not only on the type of legume but also on the pH of the soaking solution. Cooking the soaked chickpeas completely eliminated TIA, irrespective of the type of soaking solution employed (Table 4). Since compounds with TIA are heat labile, heat treatments seems to be effective in reducing the TIA. These results indicate that the cooking treatment can inactivate the action of trypsin inhibitors, potentially improving legume protein digestibility. In biological studies carried out on rats using the processed chickpeas described in this paper, Nestares et al. [41] showed that the digestive utilisation of protein, expressed as the apparent digestibility coefficient (ADC), increased compared with the protein ADC for raw chickpeas (78.7 for raw seeds and 82–84 for cooked chickpeas), showing that the effect of processing on the increase in chickpea protein digestibility can be related to the decrease in the TIA of processed seeds. Savage and Thomson [51] also reported that cooking chickpeas in water increased the net protein utilisation by between 12% and 20%, and explained this effect as a result of trypsin inhibitor removal. These results indicate that cooking can be considered as a means of obtaining legume flour with reduced TIA levels and improved nutritional quality. Although cooking seems to destroy the TIA and improve protein digestibility, results found in the literature on the effect of heat on trypsin inhibitors are controversial. Gallardo et al. [42] reported that the TIA was different under heat conditions depending on the type of legume, being heat stable in Phaseolus and heat labile in V. faba. When high molecular weight proteins are among the compounds with TIA the heat inactivation is more rapid, whereas the presence of other TIA compounds such as phenolic compounds makes them more heat resistant [52]. Sayeed and Njaa [53] reported
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that as a consequence of cooking a sharp reduction of TIA was observed in lentils and chickpeas, although it was not eliminated completely. Vidal-Valverde et al. [50] observed the total removal of TIA after cooking presoaked lentils in different solutions, whereas in faba beans TIA was totally removed only in cooked seeds after soaking in water and citric acid. Hamza et al. [46] reported that as a consequence of cooking the activity of trypsin inhibitors in chickpeas, faba beans and lentils was removed and electrophoretic bands were not found. In chickpea seeds, Sotelo et al. [54] reported that cooking caused a 57% reduction of TIA, whilst Davinder-Kaur and Hira [55] observed total TIA elimination after cooking. Table 3 shows that dry heating caused only a 27% reduction in TIA. No information has been found on the effect of dry heating on the TIA of chickpeas. In faba beans, Griffiths [56] and Kozlowska et al. [57, 58] observed the removal of TIA after dry heating, whilst Vidal-Valverde et al. [23] found 31% TIA remaining after dry heating for 15 min at 120 7C. In lentils, Sohonie and Bhanderkar [59] observed that after 30 min of dry heating the activity of trypsin inhibitors was eliminated. However, Urbano et al. [60] noted a 58% loss in the TIA of lentils after dry heating at 121 7C for 15 min and Batra et al. [49] found that dry heating for 20 min at 121 7C abolished TIA completely in lentils. The heat source also seems to play an important role in removing TIA. Hung et al. [61] demonstrated that a large decrease in TIA was obtained when lentils were submitted to microwave treatment for 2 min, and Rajkó et al. [62] designed experiments for reducing TIA in soybeans by microwave energy. Kas et al. [63] reported that TIA was removed from faba bean seeds treated with an infrared source for 40–50 s (plus 120 s of keeping time). Batra et al. [49], working with different heat sources, concluded that water boiling was the most effective heat treatment for reducing TIA. In conclusion, cooking the presoaked chickpeas in either water, acidic or basic solution seems to be an adequate treatment for obtaining a nutritious legume flour since a fair decrease in starch was observed, the largest a-galactoside reduction was produced and TIA was completely eliminated, which improved the digestive utilisation of the protein. Acknowledgements This study was supported by Commision Interministerial de Ciencia y Tecnologia ALI 96-0480, and forms part of the PhD of C. Sotomayor.
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